engleski

THE INFLUENCE OF THE BLACK MUSTARD SAMPLE PREPARATION METHOD ON CHEMICAL COMPOSITION, ANTIPROLIFERATIVE AND PROAPOPTOTIC EFFECTS

Black mustard (Brassica nigra L.) is a plant that belongs to Brassicaceae family and contains glucosinolates which in contact with the enzyme myrosinase usually decompose into isothiocyanates which give mustard its flavour [1]. The most researched degradation products of glucosinolates are isothiocyanates, which are generally known to be the most biologically active [2], but in the presence of epithiospecifier protein (ESP), epithionitriles and nitriles may be formed [3, 4]. Isothiocyanates and their metabolites help reduce the risk of developing various types of cancer such as lung, liver, esophagus, stomach, small and large intestine [5]. Degradation products of glucosinolates may exert their antitumor properties via few different mechanisms that include the immediate detoxification of carcinogens such as the prevention of the formation of carcinogen-induced DNA-adducts [6]. The aim of this study was: to determine the influence of the sample preparation method on the chemical composition of black mustard seeds volatile isolates obtained by microwave-assisted distillation (MAD), microwave hydrodiffusion and gravity (MHG), and Clevenger hydrodistillation. Also, the aim of the study was to determine the antiproliferative activity of volatile isolates against two tumor cell lines (breast cancer cell line MDA-MB-231, and bladder cancer cell line TCCSUP) by MTT assay and its proapoptotic effects determined by flow cytometry. GC-MS analysis showed that but-3-enyl isothiocyanate is a main component in Brassica nigra L. after Clevenger hydrodistillation (80.58%). After MAD and MHG, the main component in that two isolates is 4, 5-epithio-pentanenitrile (50.70%, and 59.93%). After 72 h, MHG volatile isolate from B. nigra showed high antiproliferative activity (9, 06 µg/mL) against human breast cancer cell line MDA-MB-231. The best antiproliferative activity of B. nigra MHG volatile isolate against human bladder cancer cell line TCCSUP was after 48 h and 72 h (58.95 and 27.21 µg/mL).

Brassicaceae family ; Black mustard ; Isothiocyanates ; Epithionitriles ; Antiproliferative activity

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